Noureddini Sam C, Krendelshchikov Alexander, Simonenko Vera, Hedley Susan J, Douglas Joanne T, Curiel David T, Korokhov Nikolay
VectorLogics, Inc., 550 11th Street South, Birmingham, AL 35294, USA.
Virus Res. 2006 Mar;116(1-2):185-95. doi: 10.1016/j.virusres.2005.10.002. Epub 2005 Nov 15.
The utility of adenovirus serotype 5 (Ad5)-based vectors for gene therapy applications would be improved by cell-specific targeting. However, strategies to redirect Ad5 vectors to alternate cellular receptors via replacement of the capsid fiber protein have often resulted in structurally unstable vectors. In view of this, we hypothesized that the selection of modified adenoviruses during their rescue and propagation would be a straightforward approach that guarantees the generation of functional, targeted vectors. Based on our first generation fiber-fibritin molecule, several new chimeric fibers containing variable amounts of fibritin and the Ad5 fiber shaft were analyzed via a new scheme for Ad vector selection. Our selected chimera, composed of the entire Ad5 fiber shaft fused to the 12th coiled-coil segment of fibritin, is capable of efficient capsid incorporation and ligand display. Moreover, transduction by the resultant vector is independent of the expression of the native Ad5 receptor. The incorporation of the Fc-binding domain of Staphylococcus aureus protein A at the carboxy terminus of this chimeric fiber facilitates targeting of the vector to a variety of cellular receptors by means of coupling with monoclonal antibodies. In addition, we have concluded that Ad5 vectors incorporating individual targeting ligands require individual optimization of the fiber-fibritin chimera, which may be accomplished by selecting the optimal fiber-fibritin variant at the stage of rescue of the virus in cells of interest, as described herein.
通过细胞特异性靶向可提高基于5型腺病毒（Ad5）的载体在基因治疗应用中的效用。然而，通过替换衣壳纤维蛋白将Ad5载体重定向至其他细胞受体的策略常常导致载体结构不稳定。鉴于此，我们推测在修饰腺病毒的拯救和增殖过程中进行选择将是一种直接的方法，可确保产生功能性靶向载体。基于我们的第一代纤维-纤维蛋白分子，通过一种新的腺病毒载体选择方案分析了几种含有不同量纤维蛋白和Ad5纤维杆的新型嵌合纤维。我们选择的嵌合体由与纤维蛋白的第12个卷曲螺旋结构域融合的完整Ad5纤维杆组成，能够有效地整合到衣壳并展示配体。此外，所得载体的转导不依赖于天然Ad5受体的表达。在这种嵌合纤维的羧基末端掺入金黄色葡萄球菌蛋白A的Fc结合结构域，可通过与单克隆抗体偶联促进载体靶向多种细胞受体。此外，我们得出结论，掺入单个靶向配体的Ad5载体需要对纤维-纤维蛋白嵌合体进行单独优化，这可通过在感兴趣的细胞中病毒拯救阶段选择最佳的纤维-纤维蛋白变体来实现，如本文所述。
